Making your own TAQ polymerase: Issues?

D. KIM dkim at nmsu.edu
Wed Jun 11 11:41:50 EST 1997


Hello:

I have read methods published in Nucleic Acids Research and in
BioTechniques for preparing Taq polymerase produced by recombinant E.
coli.  I would like to try this out, and the plasmid is available via
ATCC.

I do not know what are the legal issues involved in producing our own Taq
for research purposes (as a PCR reagent).  Is this illegal?  Would I have
to purchase some kind of license from Hoffman-LaRoche?  Or, could it be
done freely as long as it is not sold?

Also, it is known that DNA Pol I-type polymerases, such as Taq, have a
strong bias against ddNTPs, making them problematic for cycle sequencing.
However, a single amino acid change (switch a Phe for Tyr) will remove
this bias.  I would like to know if I would be allowed to modify the gene
for Taq polymerase by site-directed mutagenesis (and maybe remove the
5'-3' exonuclease, while I am at it) to make my own thermo-sequenase-like
enzyme?  Again, is this a legal problem?

Thank you for your answers.  I hope this generates informed discussion,
and not a lot of guessing.  I would like to have fairly authoritative
answers (with the caveat that I am not taking this as actual legal advice,
for which I would consult a licensed attorney.)

Daniel Kim, who does not want to get in more trouble than he is already
in.
dkim at nmsu.edu




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